Cytochrome c is a well-characterized mobile electron transport protein that is essential to energy conversion in all aerobic organisms. In mammalian cells, this highly conserved protein is normally localized to the mitochondrial inter-membrane space. More recent studies have identified cytosolic cytochrome c as a factor necessary for activation of apoptosis. During apoptosis, cytochrome c is trans-located from the mitochondrial membrane to the cytosol, where it is required for activation of caspase-3 (CPP32). Overexpression of Bcl-2 has been shown to prevent the translocation of cytochrome c, thereby blocking the apoptotic process. Overexpression of Bax has been shown to induce the release of cytochrome c and to induce cell death. The release of cytochrome c from the mitochondria is thought to trigger an apoptotic cascade, whereby Apaf-1 binds to Apaf-3 (caspase-9) in a cytochrome c-dependent manner, leading to caspase-9 cleavage of caspase-3. This MAb recognizes total cytochrome C which includes both apocytochrome (i.e. cytochrome in the cytosol without heme attached) and holocytochrome (i.e cytochrome in the mitochondria with heme attached). Primary antibodies are available purified, or with a selection of fluorescent CF Dyes and other labels. CF Dyes offer exceptional brightness and photostability. Note: Conjugates of blue fluorescent dyes like CF405S and CF405M are not recommended for detecting low abundance targets, because blue dyes have lower fluorescence and can give higher non-specific background than other dye colors.
Higher concentration may be required for direct detection using primary antibody conjugates than for indirect detection with secondary antibody|Immunofluorescence: 0.5-1 ug/mL|Immunohistology formalin-fixed 0.25-0.5 ug/mL|Staining of formalin-fixed tissues requires boiling tissue sections in 10 mM citrate buffer, pH 6.0, for 10-20 min followed by cooling at RT for 20 minutes|Flow Cytometry 0.5-1 ug/million cells/0.1 mL|Western blotting 0.5-1 ug/mL|Optimal dilution for a specific application should be determined by user
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